LOCUS       sequence-200963-        7957 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing synonymous tandem MS2 coat protein 
            fused to Halo. Used to label reporter mRNAs. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7957)
  AUTHORS   Horvathova I, Voigt F, Kotrys AV, Zhan Y, Artus-Revel CG, Eglinger 
            J, Stadler MB, Giorgetti L, Chao JA
  TITLE     The Dynamics of mRNA Turnover Revealed by Single-Molecule Imaging in
            Single Cells.
  JOURNAL   Mol Cell. 2017 Nov 2;68(3):615-625.e9. doi: 
            10.1016/j.molcel.2017.09.030. Epub 2017 Oct 19.
  PUBMED    29056324
REFERENCE   2  (bases 1 to 7957)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.molcel.2017.09.030"; journalName: "Mol Cell"; date: 
            "2017-11-2- 2"; volume: "68"; issue: "3"; pages: "615-625.e9"
COMMENT     Sequence Label: phage UbiC NLS HA stdMCP Halo
FEATURES             Location/Qualifiers
     source          1..7957
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    679..804
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1301..1534
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1719..1763
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2056..2173
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2216..2614
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     regulatory      2622..2631
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2634..2654
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2658..2684
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             2658..2684
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2685..2708
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3441..4325
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     misc_feature    4345..4933
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4398..4418)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4816..4827)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5008..5241
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5360..5378)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5446..5550
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5551..5619,5620..6411)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5769..5788)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6582..7170
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7071..7090
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7324..7341
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7581..7602
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7617..7622,7623..7640,7641..7647)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7655..7671
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     7660..7682
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7679..7695
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7679..7695
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7745..7764
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             join(7956..7957,1..632)
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 gaagggctaa ttcactccca aagaagacaa gatatccttg atctgtggat ctaccacaca
       61 caaggctact tccctgatta gcagaactac acaccagggc caggggtcag atatccactg
      121 acctttggat ggtgctacaa gctagtacca gttgagccag ataaggtaga agaggccaat
      181 aaaggagaga acaccagctt gttacaccct gtgagcctgc atgggatgga tgacccggag
      241 agagaagtgt tagagtggag gtttgacagc cgcctagcat ttcatcacgt ggcccgagag
      301 ctgcatccgg agtacttcaa gaactgctga tatcgagctt gctacaaggg actttccgct
      361 ggggactttc cagggaggcg tggcctgggc gggactgggg agtggcgagc cctcagatcc
      421 tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc agatctgagc
      481 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
      541 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
      601 acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg
      661 aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca
      721 agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag
      781 gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa
      841 aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca
      901 agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt
      961 agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca
     1021 ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc
     1081 aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa
     1141 gcggccggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
     1201 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
     1261 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
     1321 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
     1381 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
     1441 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
     1501 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
     1561 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
     1621 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
     1681 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
     1741 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
     1801 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
     1861 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
     1921 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
     1981 gagacagatc cattcgatta gtgaacggat ctcgacggta tcgccgaatt cacaaatggc
     2041 agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag
     2101 aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa
     2161 aattcaaaat tttcgggttt attacaggga cagcagagat ccagtttgga ctagtggcct
     2221 ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcc tcacggcgag cgctgccacg
     2281 tcagacgaag ggcgcagcga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc
     2341 cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg
     2401 gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag
     2461 tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta
     2521 tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg
     2581 ttcttgtttg tggatcgctg tgatcgtcac ttggcggccg cgccaccatg ggcccaaaaa
     2641 agaaaagaaa agttggctac ccctacgacg tgcccgacta cgccctggaa gttctgttcc
     2701 aggggcccct gttattaatt aacgcttcta actttactca gttcgttctc gtcgacaatg
     2761 gcggaactgg cgacgtgact gtcgccccaa gcaacttcgc taacgggatc gctgaatgga
     2821 tcagctctaa ctcgcgttca caggcttaca aagtaacctg tagcgttcgt cagagctctg
     2881 cgcagaatcg caaatacacc atcaaagtcg aggtgcctaa aggcgcctgg cgttcgtact
     2941 taaatatgga actaaccatt ccaattttcg ccacgaattc cgactgcgag cttattgtta
     3001 aggcaatgca aggtctccta aaagatggaa acccgattcc ctcagcaatc gcagcaaact
     3061 ccggcatcta cgccatggcc agcaacttca cccagttcgt gctggtggac aacggcggca
     3121 ccggcgacgt gaccgtggcc cccagcaact tcgccaacgg catcgccgag tggatcagca
     3181 gcaacagcag aagccaggcc tacaaggtga cctgcagcgt gagacagagc agcgcccaga
     3241 acagaaagta caccatcaag gtggaggtgc ccaagggcgc ctggagaagc tacctgaaca
     3301 tggagctgac catccccatc ttcgccacca acagcgactg cgagctgatc gtgaaggcca
     3361 tgcagggcct gctgaaggac ggcaacccca tccccagcgc catcgccgcc aacagcggca
     3421 tctacgccga cccaccggtc gaaatcggta ctggctttcc attcgacccc cattatgtgg
     3481 aagtcctggg cgagcgcatg cactacgtcg atgttggtcc gcgcgatggc acccctgtgc
     3541 tgttcctgca cggtaacccg acctcctcct acgtgtggcg caacatcatc ccgcatgttg
     3601 caccgaccca tcgctgcatt gctccagacc tgatcggtat gggcaaatcc gacaaaccag
     3661 acctgggtta tttcttcgac gaccacgtcc gcttcatgga tgccttcatc gaagccctgg
     3721 gtctggaaga ggtcgtcctg gtcattcacg actggggctc cgctctgggt ttccactggg
     3781 ccaagcgcaa tccagagcgc gtcaaaggta ttgcatttat ggagttcatc cgccctatcc
     3841 cgacctggga cgaatggcca gaatttgccc gcgagacctt ccaggccttc cgcaccaccg
     3901 acgtcggccg caagctgatc atcgatcaga acgtttttat cgagggtacg ctgccgatgg
     3961 gtgtcgtccg cccgctgact gaagtcgaga tggaccatta ccgcgagccg ttcctgaatc
     4021 ctgttgaccg cgagccactg tggcgcttcc caaacgagct gccaatcgcc ggtgagccag
     4081 cgaacatcgt cgcgctggtc gaagaataca tggactggct gcaccagtcc cctgtcccga
     4141 agctgctgtt ctggggcacc ccaggcgttc tgatcccacc ggccgaagcc gctcgcctgg
     4201 ccaaaagcct gcctaactgc aaggctgtgg acatcggccc gggtctgaat ctgctgcaag
     4261 aagacaaccc ggacctgatc ggcagcgaga tcgcgcgctg gctgtctact ctggagattt
     4321 ccggttaatg aatcgataga tcctaatcaa cctctggatt acaaaatttg tgaaagattg
     4381 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     4441 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     4501 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     4561 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     4621 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     4681 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     4741 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     4801 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     4861 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     4921 gccgcctccc cgcctgagat cctttaagac caatgactta caaggcagct gtagatctta
     4981 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag
     5041 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     5101 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     5161 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     5221 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
     5281 ttgcaaagaa atgaatatca gagagtgaga ggcccgggtt aattaaggaa agggctagat
     5341 cattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5401 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5461 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5521 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5581 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5641 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5701 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5761 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     5821 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5881 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5941 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     6001 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6061 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6121 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6181 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6241 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6301 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6361 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6421 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6481 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6541 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6601 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6661 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6721 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6781 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6841 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6901 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6961 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7021 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7081 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7141 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7201 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     7261 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     7321 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     7381 gcgcgttggc cgattcatta atgcagcaag ctcatggctg actaattttt tttatttatg
     7441 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     7501 gaggcctagg cttttgcaaa aagctccccg tggcacgaca ggtttcccga ctggaaagcg
     7561 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     7621 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     7681 ggaaacagct atgacatgat tacgaatttc acaaataaag catttttttc actgcattct
     7741 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa ctggataact
     7801 caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac tgccaattac
     7861 ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt ttaaagaaat
     7921 tgtatttgtt aaatatgtac tacaaactta gtagttg
//
